CHARACTERIZATION OF MUSCLE PFK-DEFICIENCY

磷酸果糖激酶(PFK；EC.2.7.1.11)存在于四聚体中 人类和许多脊椎动物的同工酶形式。 三个结构位点编码三个不同的亚基，M 肌肉，L(肝脏)和P(血小板)型。在人类身上，遗传的 处于纯合子状态的肌肉PFK缺乏相关 有劳力性肌病和溶血(糖原储存 疾病类型VII)，其中杂合子状态导致 临床上无症状的携带者状态。 最近，我们描述了遗传性肌肉的发生。 英国施普林格猎犬中的PFK缺乏症。奇怪的是 与人类的同类不同，这些缺陷的狗表现出 存在更严重的溶血综合征，但明显 没有肌肉疾病。这些非典型特征如下所示 是同工酶分布模式的独特性质造成的 和狗的肌肉生理学以及一种代偿 先证者对肝脏同工酶的保留或再表达。我们 现在希望更详细地研究这种动物模型 一种评价人糖酵解酶的实验方法 有用处。具体来说，我们希望进行以下工作 研究：(1)建立一个小型繁育群体 PFK缺陷的狗和最终发展成 高尿酸血症犬；(2)常规血液学和生化 研究；(3)对这些糖酵解能力的评估 狗；(4)血液中PFK同工酶的定义 细胞、培养的细胞系和固体器官；(5)代谢 肌肉的研究；(6)犬肌肉的发育研究 和红细胞；(7)免疫细胞学、组织化学和 肌肉的电子显微镜研究。技术和/或 这些研究所需的试剂有 在我们的实验室中得到了很好的证实/提供。 预计这些研究将更详细地描述 这种人类酶病的动物模型，并评估其 作为一些人的实验模型的实用性 包括溶血、代谢性肌肉疾病、 高尿酸血症和痛风并作为骨髓的接受者 移植以及酶和基因替代疗法。

Phosphofructokinase (PFK; EC.2.7.1.11) exists in tetrameric isozymic forms in man and a number of vertebrate species. Three structural loci encode three distinct subunits, M (muscles, L (liver) and P (platelet) types. In man, inherited deficiency of muscle PFK in the homozygous state is associated with exertional myopathy and hemolysis (glycogen storage disease type VII) where as the heterozygous state results in a clinically silent carrier state. Recently, we have described the occurrence of inherited muscle PFK deficiency among English Springer Spaniel dogs. Curiously unlike their human counterparts, the deficient dogs exhibit the presence of a more severe hemolytic syndrome but an apparent absence of muscle disease. These atypical features are shown to result from a unique nature of isozyme distribution pattern and muscle physiology in the dog as well as a compensating retention or reexpression of liver isozyme by the probands. We now wish to investigate in greater detail this animal model of a human glycolytic enzymopathy to assess its experimental usefulness. Specifically, we wish to undertake the following studies: (1) Establishment of a small breeding colony of PFK-deficient dogs and eventually development of a hyperuricemic dog; (2) Routine hematological and biochemical studies; (3) Assessment of the glycolytic competence of these dogs; (4) Definition of the PFK isozymic profile of blood cells, cultured cell lines, and solid organs; (5) Metabolic studies of muscle; (6) Developmental studies of canine muscle and erythrocytes; (7) Immunocytologic, histochemical and electron microscopic studies of muscle. The techniques and/or reagents necessary for these studies are well-established/available in our laboratories. These studies are expected to characterize in greater detail this animal model of a human enzymopathy and assess its usefulness as an experimental model for a number of human diseases including hemolysis, metabolic muscle disease, hyperuricemia and gout and as a recipient of bone marrow transplantation and enzyme and gene replacement therapies.